Neuron, Vo|.

37, 577-582, February 20, 2003, Copyright 2003 by Ce|| Press

Report Drugs of Abuse and Stress Tr|gger
a Common Synapt|c Adaptat|on
|n Dopam|ne Neurons
is the sub[ect of debate (Berridge and Robinson, 1998},
there is no doubt that the discovery of this common
action of addictive drugs, despite their very different
mo|ecu|ar actions, has had a ma[or inf|uence on thinking
about both the neurobio|ogica| basis of addiction as
Dan|e| Saa|,
1,3
Yan Dong,
1,3
Antone||o Bonc|,
2
and Robert C. Ma|enka
1,
*
1
Nancy Pritzker Laboratory
Department of Psychiatry and Behaviora| Sciences
Stanford University Schoo| of Medicine
Pa|o A|to, Ca|ifornia 94304 we|| as the brain mechanisms mediating reinforcement
and reward-based |earning (Wae|ti et a|., 2001}.
2
Ernest Ga||o C|inic and Research Center
Department of Neuro|ogy and Recent|y we found that a sing|e in vivo exposure to
cocaine caused an enhancement of synaptic strength Whee|er Center for the Neurobio|ogy of Addiction
University of Ca|ifornia, San Francisco at excitatory synapses onmidbrain DAneurons(Ung|ess
et a|., 2001} and that this shared severa| features with San Francisco, Ca|ifornia 94110
|ong-term potentiation (LTP}, the |eading mode| for the
ce||u|ar changes that mediate |earning and memory
(Martin et a|., 2000}. Here we examine whether in vivo Summary
administration of other common|y abused drugs causes
a simi|ar synaptic change. Furthermore, because in both Drug seek|ng and drug se|f-adm|n|strat|on |n both an|-
ma|s and humans can be tr|ggered by drugs of abuse humans and anima|s stress faci|itates the initia| acquisi-
tion and maintenance of drug se|f-administration and themse|ves or by stressfu| events. Here, we demon-
strate that |n v|vo adm|n|strat|on of drugs of abuse can e|icit re|apse (i.e., reinstatement of drug taking after
abstinence} (Piazza and Le Moa|, 1998, Shaham et a|., w|th d|fferent mo|ecu|ar mechan|sms of act|on as we||
as acute stress both |ncrease strength at exc|tatory 2000}, we asked whether an acute stress a|so causes a
change in excitatory synaptic strength on midbrain DA synapses on m|dbra|n dopam|ne neurons. Psychoac-
t|ve drugs w|th m|n|ma| abuse potent|a| do not cause neurons ana|ogous to that caused by cocaine. Our find-
ings strengthen the idea that mechanisms invo|ved in th|s change. The synapt|c effects of stress, but not of
coca|ne, are b|ocked by the g|ucocort|co|d receptor adaptive forms of experience-dependent p|asticity such
as |earning may be patho|ogica||y usurped and contrib- antagon|st RU486. These resu|ts suggest that p|ast|c-
|ty at exc|tatory synapses on dopam|ne neurons may ute to the deve|opment of addiction (Hyman and Ma-
|enka, 2001, Nest|er, 2001}. be a key neura| adaptat|on contr|but|ng to add|ct|on
and |ts |nteract|ons w|th stress and thus may be an
attract|ve therapeut|c target for reduc|ng the r|sk of
Resu|ts
add|ct|on.
The experimenta| approach was essentia||y identica| to
Introduct|on
that used previous|y (Ung|ess et a|., 2001}. Twenty-four
hours after in vivo administration of the drug (or stress},
Understanding the neurobio|ogica| basis of addiction
s|ices containing the VTA were prepared and who|e-ce||
has important imp|ications not on|y for the treatment
recordings were obtained from DA ce||s, identified by
and prevention of this devastating and common i||ness
the presence of |arge /
h
currents (Figure 1A}. To assess
but a|so as a mode| for how experiences modify neura|
synaptic strength, the ratio of AMPA receptor-mediated
circuitry and thereby behavior. Because humans abuse
synaptic currents (AMPAR EPSCs} to NMDA receptor-
and become addicted to a wide array of |ega| (e.g.,
mediated synaptic currents (NMDAREPSCs} was ca|cu-
nicotine, ethano|} and i||icit (e.g., cocaine, heroin} drugs,
|ated (Figure 1B}. This measure offers the important ad-
a va|uab|e strategy has been to |ook for adaptations in
vantage that it is independent of the number of synapses
the brain that are common to different c|asses of ad-
activated and therefore independent of such variab|es
dictive substances. Such an approach |ed to the rea|iza-
as the positioning of the e|ectrodes or anatomy of the
tionthat onecritica||yimportant component of the neura|
tissue. We first retested the effects of cocaine and com-
circuitsmediatingaddictionisthemeso|imbic dopamine
pared its effects to those of amphetamine, a psycho-
(DA} system consisting of midbrain DA ce||s, |ocated
stimu|ant that shares some mechanistic features with
primari|y in the ventra| tegmenta| area (VTA} and the
cocaine in that it too enhances brain DA |eve|s. Both
nuc|eus accumbens (NAc}, which receives a dense pro-
psychostimu|ant drugs caused a simi|ar increase in the
[ection from the VTA (Everitt and Wo|f, 2002, Ke||ey and
ratio of AMPAR to NMDAR EPSCs (Figure 1C} (sa|ine,
Berridge, 2002, Koob et a|., 1998, Nest|er, 2001, Wise,
0.42 .09, n 14 ce||s, cocaine, 0.70 .08, n 8,
1996}. A|| drugs of abuse appear to act direct|y in the
amphetamine, 0.70 .09, n 12}, indicating that |ike
VTA and/or NAc to cause increases in DA |eve|s in the
cocaine, in vivo administration of amphetamine causes
NAc (Koob et a|., 1998, Nest|er, 2001, Wise, 1996}. A|-
an increase in excitatory synaptic transmission onto DA
though whether this is sufficient or even abso|ute|y nec-
neurons.
essary for the reinforcing properties of drugs of abuse
Is this in vivo drug-induced synaptic p|asticity in mid-
brain DA neurons |imited to psychostimu|ants or does
it a|so occur in response to other addictive substances?
*Correspondence. ma|enka@stanford.edu
3
These authors contributed equa||y to this work. Wetestedthree other common|y abuseddrugsthat have
Neuron
578
Figure 1. Psychostimu|ant Drugs Enhance the AMPA/NMDA EPSC Ratio at Excitatory Synapses on Midbrain DA Ce||s
(A} Examp|e of /h currents used to identify midbrain DA ce||s (ca|ibration bars. 20 pA/50 ms}.
(B} Top traces show examp|e from a contro| ce|| of how AMPA/NMDA ratios were obtained. EPSCs were recorded at 40 mV (Dua| trace},
and then D-APV (50 M} was app|ied to obtain the AMPA EPSC. The NMDA EPSC was obtained by digita| subtraction of the AMPA EPSC
from the dua| EPSC. Bottom traces show examp|es of AMPA and NMDA EPSCs obtained from cocaine- and amphetamine-treated anima|s
(ca|ibration bars. 20 pA/15 ms}.
(C} Summary of AMPA/NMDA ratios obtained from anima|s administered sa|ine, cocaine, or amphetamine (asterisk indicates p 0.02}. In this
and a|| subsequent figures, numbers within the bars indicate the number of ce||s examined.
very different mo|ecu|ar mechanisms of action. (1} mor- tor common|y used in the treatment of depression, nor
carbamazepine, which is a common treatment for sei- phine, which acts on opioid receptors, (2} nicotine,
which acts on brain nicotinic receptors, and (3} ethano|, zure disorders and bipo|ar disorder (i.e., manic-
depressive i||ness} (Nest|er et a|., 2001}, had an effect which acts on a number of different neurotransmitter
receptors (Koob et a|., 1998, Nest|er, 2001}. In vivo ad- on the ratio of AMPAR to NMDAR EPSCs (Figure 3}
(sa|ine, 0.44 .02, n 14 ce||s, f|uoxetine, 0.44 .05, ministration of a|| three of these substances caused
significant increases in the ratio of AMPAR to NMDAR n 9, carbamazepine, 0.41 .05, n 8}. These resu|ts
provide further support for the hypothesis that changes EPSCs measured 24 hr |ater (Figure 2} (sa|ine, 0.46
.02, n 22 ce||s, morphine, 0.68 .04, n 9, nicotine, in strength at excitatory synapses on midbrain DA ce||s
may contribute to the addictive properties of mu|tip|e 0.67 .07, n 14, ethano|, 0.64 .06, n 10}.
Thus far we have demonstrated that five different c|asses of abused substances.
The most difficu|t aspect of addiction treatment is the abused drugs (cocaine, amphetamine, morphine, nico-
tine, and ethano|} a|| cause a simi|ar enhancement at |ong-|asting risk of re|apse (O'Brien, 1997}, individua|s
can be abstinent for months or even years but sti|| are excitatory synapses on midbrain DA ce||s. These are
surprising resu|ts since these substances have very dif- susceptib|e to cravings that can stimu|ate renewed drug
seeking and taking. In both humans and anima|s, rein- ferent mo|ecu|ar mechanisms of action. They a|so raise
the important question of whether this modification has statement of drug seeking and se|f-administration can
be triggered by sing|e doses of drug, stimu|i previous|y something to do with the addictive potentia| of these
substances. A|ternative|y, it might simp|y be the conse- associated with drug taking, or stressfu| events (Piazza
and Le Moa|, 1998, Robinson and Berridge, 2000, Sha- quence of administering any type of psychoactive sub-
stance. To address this issue, we asked whether admin- ham et a|., 2000}. Stress a|so can faci|itate the initia|
acquisition of drug se|f-administration, perhaps by en- istration of common|y used therapeutic drugs, which
are known to have profound effects on emotiona| states hancing the reinforcing efficacy of drugs of abuse (Pi-
azza and Le Moa|, 1998}. We therefore tested whether in humans but no addictive |iabi|ity, a|so caused a
change at excitatory synapses on midbrain DA ce||s. anacute stress wou|d affect excitatorysynapticstrength
on midbrain DAce||s. Anima|s were sub[ect to a modified Neither f|uoxetine, a specific serotonin-reuptake inhibi-
Figure 2. Common|y Abused Drugs Other
than Psychostimu|ants a|so Increase the
AMPA/NMDA Ratio
(A} Examp|es of AMPA and NMDA EPSCs ob-
tained from anima|s given the indicated sub-
stance (ca|ibration bars. 20 pA/15 ms}.
(B} Summary of AMPA/NMDA ratios obtained
from anima|s administered sa|ine, morphine,
nicotine, or ethano| (asterisk indicates p
0.03}.
Drugs and Stress Cause Synaptic Changes in the VTA
579
(Figures 4A and 4B} (stress p|us RU486, 0.38 .04, n
9 ce||s}, imp|ying a critica| ro|e for GR activation. The
effectiveness of RU486 a|so a||owed us to address the
question of whether the synaptic effect of cocaine is
due to an unrecognized drug-induced stress reaction
rather than its reinforcing and/or addictive property.
However, RU486 did not prevent the increase in the
AMPARtoNMDAREPSCratio e|icitedby in vivo cocaine
administration (Figure 4C} (sa|ine, 0.45 .03, n 14
ce||s, cocaine, 0.61 .13, n 7, cocaine p|us RU486,
0.69 .12, n 9}. This indicates that the synaptic conse-
quences of in vivo cocaine exposure do not require GR
activation and therefore are un|ike|y to be due to an
acute stress response.
Figure 3. Common|y Used Psychoactive but Nonaddictive Drugs
Do Not Affect the AMPA/NMDA Ratio
(A} Examp|es of AMPA and NMDA EPSCs obtained from anima|s
D|scuss|on
given the indicated substance (ca|ibration bars. 20 pA/15 ms}.
(B} Summary of AMPA/NMDAratios obtained fromanima|s adminis-
We have demonstrated that in vivo administration of five
tered sa|ine, f|uoxetine, or carbamazepine.
different drugs of abuse with very different mo|ecu|ar
mechanisms of action a|| e|icit an enhancement of
strength at excitatory synapses on midbrain DA neu- Porso|t forced swimtask, a common|y used acute stres-
sor (Huber et a|., 2001}, and s|ices were prepared 1 day rons. Two psychoactive but therapeutic and nonad-
dictive medications did not cause such a change. This |ater. This resu|ted in an increase in the ratio of AMPAR
to NMDAREPSCs even |arger than that e|icited by drugs degree of specificity suggests that this in vivo, drug-
induced synaptic p|asticity in DA neurons may be one of abuse (Figures 4A and 4B} (sa|ine, 0.46 .03, n 11
ce||s, stress p|us sa|ine, 0.82 .04, n 24}. To begin important component of the neura| circuit adaptations
that contribute to core features of addiction. Because to exp|ore whether this stress-induced synaptic modifi-
cation shares mechanisms with the cocaine-e|icited syn- of theprofoundeffect of stressin faci|itatingthe initiation
and reinstatement of drug se|f-administration, we a|so aptic enhancement, we administered the NMDAR antago-
nist MK-801 prior to the co|d water swim. This stress examined whether an acute stress might have an effect
simi|ar to that of drugs of abuse and found that it too protoco| b|ocked the increase in the ratio of AMPAR to
NMDAR EPSCs (Figures 4A and 4B} (stress p|us MK801, causeda |arge increase in synaptic strength at this same
popu|ation of excitatory synapses. Based on these ob- 0.51 .06, n 15}, indicating that, |ike the effects of
cocaine (Ung|ess et a|., 2001}, the synaptic changes due servations, we suggest that this enhancement of strength
at excitatory synapses on midbrain DA neurons a|so is a to the acute stress required activation of NMDARs.
One of the ma[or consequences of acute stress is key neura| adaptation that contributes to the effects of
stress on drug seeking and re|apse. increased secretion of corticosteroids and activation
of g|ucocorticoid receptors (GRs}. To test whether GR The conc|usion that the increases in the AMPAR to
NMDAREPSCratio e|icitedby drugs of abuse andstress activation p|ayed a ro|e in mediating the effects of stress
on synaptic strength in midbrain DAneurons, we admin- ref|ect an enhancement of AMPAR-mediated excitatory
synaptic transmission is based on previous work that ex- istered the GRantagonist RU486 (Cadepondet a|., 1997,
Marine||i et a|., 1998} prior to p|acing anima|s in the co|d amined the mechanisms under|ying the cocaine-induced
increase in this ratio (Ung|ess et a|., 2001}. We cannot water. This comp|ete|y b|ocked the synaptic change
Figure 4. Acute Stress Increases the AMPA/NMDA Ratio
(A} Examp|es of AMPA and NMDA EPSCs obtained from a contro| anima| and anima|s experiencing stress p|us the indicated substance
(ca|ibration bars. 20 pA/15 ms}.
(B} Summary of AMPA/NMDA ratios obtained from anima|s administered sa|ine or experiencing stress a|ong with administration of sa|ine, the
NMDA receptor antagonist MK801, or the g|ucocorticoid receptor antagonist RU486 (asterisk indicates p 0.01}.
(C} Summary of AMPA/NMDA ratios obtained from anima|s administered sa|ine, cocaine, or cocaine p|us RU486 (asterisk indicates p 0.05}.
Neuron
580
ru|e out, however, additiona| mechanisms that cou|d with DA ce|| firing are granted high appetitive or motiva-
contribute to this increase in the AMPAR to NMDAR tiona| significance. We suggest that by increasing syn-
EPSC ratio. A decrease in the magnitude of the NMDAR aptic strength at excitatory synapses on midbrain DA
EPSC due to changes in NMDAR function and/or num- ce||s, drugs of abuse or stress enhance the motivationa|
ber c|ear|y wou|d increase the ratio, as wou|d decreases significance of drugs themse|ves as we|| as stimu|i
in the NMDAR EPSC due to the extent of g|utamate c|ose|y associated with drug seeking and se|f-adminis-
spi||over (Ku||mann, 2000} shou|d spi||over occur nor- tration.
ma||y in this brain region. In previous work an attempt
Consistent with this hypothesis, there is considerab|e
was made to address the first of these possibi|ities by
evidence that stress can enhance the rewarding efficacy
examining the inward currents generated by app|ication
of drugs, |ike|y via inf|uences on the meso|imbic DA
of NMDA (Ung|ess et a|., 2001}. However, this manipu|a-
system (Piazza and Le Moa|, 1998}. Indeed, stress, |ike
tion activates extrasynaptic NMDARs and thus negative
drugs of abuse, causes an increase in DA |eve|s in the
resu|ts are not conc|usive. Another possibi|ity is that
NAc and prefronta| cortex (Horger and Roth, 1996, Pi-
if norma||y synaptic AMPARs on midbrain DA neurons
azza and Le Moa|, 1998}. Stress has a|so been reported
exhibit inward rectification (i.e., pass more current in
to affect synaptic p|asticity in the hippocampus. Specifi-
the inward than in the outward direction}, a drug- or
ca||y, acute stress appears to rapid|y inhibit the genera-
stress-induced decrease in the degree of inward rectifi-
tion of LTP and faci|itate the generation of LTD (Shors
cation wou|d resu|t in an increase in the AMPAR to
et a|., 1989, Kim et a|., 1996, Xu et a|., 1997, 1998}.
NMDAR EPSC ratio when measured at 40 mV (since
Whether this invo|ves an increase in basa| synaptic
the modifiedAMPARs wi|| nowpass more current at 40
strength or rather some modification of the triggering
mV a|though the same current at hyperpo|arized mem-
mechanisms for synaptic p|asticity is debated (Kim et
brane potentia|s}. However, such a change in the bio-
a|., 1996, Xu et a|., 1997}.
physica| properties of AMPARs cannot account for our
One important unanswered question is whether a|| or
resu|ts since a c|ear increase in the AMPAR to NMDAR
critica| subsets of excitatory afferent inputs to midbrain
EPSC ratio in response to cocaine administration was
DA ce||s are strengthened by in vivo drugs or stress.
detected when AMPAR EPSCs were measured at 70
Strengthening of prefronta| cortica| inputs may be par-
mV rather than 40 mV (unpub|ished observations}.
ticu|ar|y important, as this has been suggested to p|ay
There are severa| corre|ative findings that can be
a critica| ro|e in the deve|opment of behaviora| sensitiza-
viewed as being consistent with the hypothesis that
tion, a prominent mode| for core features of addiction
drugs of abuse enhance AMPAR-mediated synaptic
inc|uding craving (Everitt and Wo|f, 2002, Robinson and
transmission in midbrain DAce||s. First, repeatedadmin-
Berridge, 2000}. On the other hand, modifications of the
istration of cocaine or amphetamine caused an increase
recent|y described input from the bed nuc|eus of the
in the sing|e unit responses of VTA DA ce||s to AMPA
stria termina|is (Georges and Aston-Jones, 2002} may
but not to NMDA, an effect that was observed 3 but not
be particu|ar|y important for mediating the effects of
14 days after cessation of drug administration (Zhang
stress (Shahamet a|., 2000}. Whatever sets of synapses
et a|., 1997}. Second, both repeated administration of
are affected, by demonstrating a synaptic modification
cocaine or morphine as we|| as repeated restraint stress
common|y e|icited by addictive drugs and stress, we
increased |eve|s of the AMPAR subunit G|uR1 in the VTA
provide a neura| mechanism that may contribute to the
(as we|| as the NMDAR subunit NR1} (Fitzgera|d et a|.,
c|inica||y important interactions between addiction and
1996, but see Lu et a|., 2002}. Third, vira|-mediated over-
stress as we|| as one potentia||y important target for
expression of G|uR1 in the VTA enhanced the |ocomotor
therapeutic interventions in treatment of addictive dis-
stimu|ating and rewarding properties of morphine (Car-
orders.
|ezon et a|., 1997}. These sorts of findings have |ed to
a hypothesis that e|evated |eve|s of G|uR1 in the VTA
are one primary trigger |eading to the comp|ex cascade
Exper|menta| Procedures
of mo|ecu|ar and circuit adaptations that under|ie sensi-
tization to drugs of abuse (Car|ezon and Nest|er, 2002}.
An|ma|s and In V|vo Man|pu|at|ons
Accepting that drugs of abuse and stress do enhance C57/B|6 mice (21- to 30-days-o|d} were used for a|| experiments.
For drug administration, anima|s were in[ected i.p. at the same time the strength of excitatory afferent inputs to midbrain DA
each day with sa|ine (vo|ume matched for experimenta| in[ections},
ce||s, how might this contribute to the deve|opment of
cocaine (15 mg/kg}, d-amphetamine (10 mg/kg}, morphine (10 mg/kg},
addiction and/or re|apse? There are three genera| hypoth-
nicotine (0.5 mg/kg}, ethano| (20 mg/kg}, f|uoxetine (10 mg/kg}, or
eses concerning the functions of midbrain DA ce||s and
carbamazepine (15 mg/kg}. Drugs were obtained from Sigma (ex-
their pro[ections to the NAc. DAce|| firing and the conse-
cept for ethano|, which was obtained from the Stanford Hospita|
quent re|ease of DA in the NAc may. (1} signify reward
pharmacy}. Acute stress was e|icited with a modified Porso|t forced
(Wise, 1996} or changes in the positive incentive va|ue
swim task. Anima|s were p|aced in a 1 | graduated cy|inder con-
of stimu|i (Koob, 1996}, (2} serve as a reward prediction taining 400 m| of co|d (6 C} water for 4-6 min. MK801 (1 mg/kg in
sa|ine} or RU486 (40 mg/kg in DMSO} was administered 15-30 min signa| and thereby p|ay an important ro|e in reward-
prior to the co|d water swim. RU486 was administered 30 min prior
based |earning (Wae|ti et a|., 2001}, or (3} be invo|ved in
to cocaine administration for the experiments i||ustrated in Figure
signa|ing "incentive sa|ience," which in the context of
4C. The dose of RU486 was chosen based on pub|ished reports
addiction suggests that adaptations in the meso|imbic
showing variab|e efficacy of 20-25 mg/kg (Pieretti et a|., 1991,
DA system |ead to "wanting" (i.e., craving} drugs a|-
Douma et a|., 1998, Xu et a|., 1998} and the concern that because
though not necessari|y "|iking" them(Robinson and Ber-
rodents |ack a specific binding protein for RU486, brain concentra-
ridge, 2000}. These non-mutua||y exc|usive hypotheses
tions of RU486 fo||owing periphera| administration are |ow (Heikin-
heimo and Kekkonen, 1993}. share the featurethat externa| stimu|i that are associated
Drugs and Stress Cause Synaptic Changes in the VTA
581
E|ectrophys|o|ogy pristone}. mechanisms of action and c|inica| uses. Annu. Rev. Med.
48, 129-156. Anima|s were anesthetized with ha|othane and sacrificed 24-30 hr
after the in vivo manipu|ation. A|| remaining procedures were essen-
Cameron, D.L., Wessendorf, M.W., and Wi||iams, J.T. (1997}. A sub-
tia||y as described previous|y (Ung|ess et a|., 2001}. Brief|y, a b|ock
set of ventra| tegmenta| area neurons is inhibited by dopamine,
of tissue containing midbrain was s|iced in the horizonta| p|ane (250
5-hydroxytryptamine and opioids. Neuroscience 77, 155-166.
m} in ice-co|d |ow Ca
2
so|ution (containing in mM. 126 NaC|,
Car|ezon, W.A., and Nest|er, E.J. (2002}. E|evated |eve|s of G|uR1 in
1.6 KC|, 1.2 NaH
2
PO
4
, 1.2 MgC|
2
, 0.625 CaC|
2
, 18 NaHCO
3
, and 11
the midbrain. a trigger for sensitization to drugs of abuse? Trends
g|ucose}. S|ices were transferred to a ho|ding chamber containing
Neurosci. 25, 610-615.
artificia| cerebrospina| f|uid (ACSF, in mM. 126 NaC|, 1.6 KC|, 1.2
Car|ezon, W.A., Boundy, V.A., Hai|e, C.N., Lane, S.B., Ka|b, R.G.,
NaH2PO4, 1.2 MgC|2, 2.5 CaC|2, 18 NaHCO3, and 11 g|ucose} and
Neve, R.L., and Nest|er, E.J. (1997}. Sensitization to morphine in-
equi|ibriated at 31 C -34 C for at |east 1 hr. Picrotoxin (100 M} was
duced by vira|-mediated gene transfer. Science 277, 812-814.
added to the ACSF for recording, to b|ock GABA
A
receptor-mediated
inhibitory postsynaptic potentia|s. A|| so|utions were bubb|ed with Douma, B.R.K., Korte, S.M., Buwa|da, B., |a F|eur, S.E., Bohus, B.,
95¾ O2/5¾ CO2 and perfused over the s|ice at a rate of 2.5 m|/min. and Luiten, P.G.M. (1998}. Repeated b|ockade of minera|ocorticoid
Ce||s were visua|ized with an upright microscope using infrared receptors, but not of g|ucocorticoid receptors impairs food re-
i||umination, and who|e-ce|| vo|tage-c|amp recordings were made warded spatia| |earning. Psychoneuroendocrino|ogy 23, 33-44.
with an Axopatch 1D amp|ifier (Axon Instruments}. E|ectrodes (2-6
Everitt, B.J., and Wo|f, M.E. (2002}. Psychomotor stimu|ant addic-
M } contained in mM. 117 cesium methansu|fonic acid, 20 HEPES,
tion. a neura| systems perspective. J. Neurosci. 22, 3312-3320.
0.4 EGTA, 2.8 NaC|, 5 TEA-C|, 2.5 MgATP, 0.25 MgGTP (pH 7.2-7.4},
Fitzgera|d, L.W., Ortiz, J., Hamedani, A.G., and Nest|er, E.J. (1996}.
275-285 mOsm. Dopamine ce||s were identified by the presence of
Drugs of abuse and stress increase the expression of G|uR1 and
a |arge /
h
current, which was evoked by ho|ding ce||s at 70 mV
NMDAR1 g|utamate receptor subunits in the rat ventra| tegmenta|
and stepping to 120 mV in 10 mV increments. Whi|e /
h
is present
area. common adaptations among cross-sensitizing agents. J. Neu-
in 90¾ of dopamine neurons (Cameron et a|., 1997, Neuhoff et
rosci. 16, 274-282.
a|., 2002}, we recognize that its presence does not unequivoca||y
Georges, F., and Aston-Jones, G. (2002}. Activation of ventra| teg-
identify dopamine ce||s in midbrain s|ices. However, both in previous
menta| area ce||s by the bed nuc|eus of the stria termina|is. a nove|
work (Ung|ess et a|., 2001} and in the present experiments, this
excitatory amino acid input to midbrain dopamine neurons. J. Neu-
criterion was sufficient to obtain c|ear differences between contro|
rosci. 22, 5173-5187.
and experimenta| ce||s.
A bipo|ar stimu|ating e|ectrode was p|aced 100-300 m rostra| to Heikinheimo, O., and Kekkonen, R. (1993}. Dose-response re|ation-
the recording e|ectrode and was used to stimu|ate excitatory affer- ships of RU 486. Ann. Med. 25, 71-76.
ents at 0.1 Hz. Ce||s were initia||y he|d at 70 mV for 5-15 min to
Horger, B.A., and Roth, R.H. (1996}. The ro|e of mesoprefronta| dopa-
ensure the stabi|ity of EPSCs and then were depo|arized to 40
mine neurons in stress. Crit. Rev. Neurobio|. 10, 395-418.
mV. Ce||s were again monitored for 5-15 min, at which point D-APV
Huber, J.D., Dar|ing, S.F., Park, K.K., and So|iman, K.F. (2001}. The
(50 M} was app|iedfor 10-20 min. To ca|cu|ate the AMPAR/NMDAR
ro|e of NMDA receptors in neonata| cocaine-induced neurotoxicity.
ratio, a contro| dua| component EPSC was obtained by averaging
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20-30 consecutive responses immediate|y before app|ication of
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Acknow|edgments
26, 1-13.
Marine||i, M., Aouizerate, B., Barrot, M., Le Moa|, M., and Piazza,
This work was supported by grants from NIDA (R.C.M. and A.B.},
P.V. (1998}. Dopamine-dependent responses to morphine depend
the Zaffaroni Innovation Fund for Addiction Research (R.C.M.}, and
on g|ucocorticoid receptors. Proc. Nat|. Acad. Sci. USA 95, 7742-
Department of the Army (Award No. DAMD17-01-10736}, U.S. Army
7747.
Medica| Research Acquisition Activity (Fort Detrick, MD} (A.B.}. D.S.
Martin, S.J., Grimwood, P.D., and Morris, R.G. (2000}. Synaptic p|as-
was supported by a Pfizer Postdoctora| Fe||owship. We thank Mark
ticity and memory. an eva|uation of the hypothesis. Annu. Rev. Neu-
Ung|ess and David Lyons for technica| assistance and discussions.
rosci. 23, 649-711.
Nest|er, E.J. (2001}. Mo|ecu|ar basis of |ong-term p|asticity under|y-
Received. October 22, 2002
ing addiction. Nat. Rev. Neurosci. 2, 119-128. Revised. December 17, 2002
Nest|er, E.J., Hyman, S.E., and Ma|enka, R.C. (2001}. Mo|ecu|ar Neu-
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